Application of lactobacillus johnsonii in preventing and/or treating inflammatory bowel diseases
A technology for Lactobacillus johnsonii and inflammatory bowel disease, applied in the field of oral microecological preparations, can solve problems such as insignificant curative effect, and achieve the effect of improving symptoms
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Embodiment 1
[0060] Example 1. Isolation, identification and cultivation of Lactobacillus johnsonii
[0061] 1.1. Isolation of Lactobacillus johnsonii
[0062] Place mouse colonic digesta in a 1.5ml centrifuge tube, add 500μl PBS, and vortex for 1 minute to make it completely suspended. The obtained suspension was diluted with 10-fold serial gradient, spread on MRS (Man, Rogosa and Sharpe) agar plate, and incubated overnight at 37°C.
[0063] 1.2. Identification of Lactobacillus johnsonii
[0064] Pick the single clone of Lactobacillus johnsonii in the MRS liquid medium, amplify the full length of 16S rDNA after static culture, and the primer sequences are respectively:
[0065] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3';
[0066] 1492R: 5'-CGGYTACCTTGTTACGACTT-3'.
[0067] The PCR reaction system is shown in Table 1:
[0068] Table 1. PCR reaction system
[0069]
[0070] The PCR reaction conditions are as follows:
[0071]
[0072] The obtained PCR products were detected by 1% agarose ...
Embodiment 2
[0076] Example 2. Prevention and treatment of Lactobacillus johnsonii on mice with inflammatory bowel disease
[0077] 2.1. Cultivation of Citrobacter
[0078] Citrobacter (DBS100) was activated twice and then inoculated into an Erlenmeyer flask containing 50mL of LB (Luria-Bertani) medium, sealed to form a micro-aerobic environment, cultured in a constant temperature shaker at 37°C, 220rpm for 12 hours, and set aside.
[0079] 2.2. Establishment, grouping and administration of inflammatory bowel disease mouse model
[0080] After a one-week adaptation period, the mice were randomly divided into 4 groups according to body weight, 12 in each group, including:
[0081] Control group: mice in this group were fed with PBS every 2 days;
[0082] Illness group (i.e. Citrobacter gavage group): mice in this group were fed with PBS once every 2 days, and on the 8th day were fed with 2×10 9 CFU of Citrobacter, and then continued to feed for a week, wherein PBS was fed once every 2 da...
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